The overall objectives of this application are to conduct research aimed at improving the treatment of bladder cancer by developing methods to assess the risk of progression and monitor response to therapy in superficial and invasive bladder cancer. It also aims at developing methods for sensitive non-invasive detection of bladder cancer to be used in early detection and in the monitoring of patients with superficial disease to decrease the morbidity involved with the monitoring of recurrence. Cooperative clinical studies to evaluate promising diagnostic and prognostic markers have been undertaken and will be pursued with other members of the Network. We will continue to share clinical material, unique tumor markers, access to a large cohort of primary Ta,T1 tumors and expertise in clinical studies and in immunological and molecular biology test development. The specific aims of this translational research program will be: AIM 1 will be to evaluate cytological markers for the detection and monitoring of bladder tumors. This will involve completing the phase III network study on exfoliated cells and attempting to develop other immunocytological assays for the diagnosis and monitoring of bladder cancer. AIM 2 will be to pursue the development of a rapid and sensitive urine test for bladder cancer based on capture assays using a panel of mAbs. AIM 3 will involve the development of new methods and reagents for evaluating the prognostic and the metastatic potential of invasive bladder cancers. We will pay particular attention to obtain reagents capable of detecting in formalin-fixed tissues the T138 antigen, a promising marker of tumor progression which appears to be complementary to p53. We will also develop a RT-PCR based blood test assay for staging of bladder cancer using probes to the cDNA of the urothelial antigen gp54 which we cloned recently. The test will also involve the capture of epithelial cells in the blood by immunobeads coated with mAbs to gp54. AIM 4 will be the evaluation of the molecular markers mdr1, MRP, MT, MTx receptor and Bcl-2 on tumor aspirates enriched by gp54 immunobeads capture to monitor response to systemic M-VAC chemotherapy. A similar approach will be used to test to test for expression of the BCG resistance gene monocytes and epithelial cells of bladder tumors in patients undergoing intravesical BCG treatment. AIM 5 will be to pursue our analysis of genetic anomalies in the most common primary Ta,T1 tumors, taking advantage of our well-characterized bank which represents an ideal source of material for identification of early events in bladder carcinogenesis. The initial focus will be on detailed analysis of chromosome 9 by microsatellite and by standard RFLP for the p16 and p15 genes which are suspected to be frequently altered in Ta tumors. Presence of these anomalies will be correlated with presence of HPV DNA, p53 mutations and the MDM2 oncogene.